Khademi Farzad, Sahebkar Amirhossein, Fasihi-Ramandi Mahdi, Taheri Ramezan Ali
Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran.
Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
APMIS. 2018 Jun;126(6):509-514. doi: 10.1111/apm.12851.
A promising strategy for preventing illness and death caused by Mycobacterium tuberculosis (Mtb) is vaccination. In this study, we aimed to evaluate the capacity of a multicomponent vaccine comprising HspX/EsxS-fused protein, PLGA (poly (lactide-co-glycolide)) and DOTAP (1, 2-dioleoyl-3-trimethylammonium propane) in eliciting immune responses against Mtb in BALB/c mice. A preparation of PLGA nanoparticles (NPs) containing fused protein and DOTAP adjuvant was made using double emulsion solvent evaporation (w/o/w) and lipid film hydration methods, respectively. After three subcutaneous immunization of BALB/c mice with various formulations, ELISA technique was used to measure interferon-γ (IFN-γ) and interleukin-4 (IL-4) cytokines levels in splenocytes as well as serum anti-HspX/EsxS IgG1 and IgG2a titers. The results of the current study showed that PLGA/HspX/EsxS/DOTAP formulation was able to induce higher levels of FN-γ, IgG1, and IgG2a responses compared with BCG as the positive control, HspX/EsxS, HspX/EsxS/DOTAP and PLGA/HspX/EsxS formulations. Our results suggest that PLGA NPs, as delivery system, and DOTAP, as adjuvant, have a good potential to enhance immune responses against HspX/EsxS antigen after subcutaneous immunization of BALB/c mice.
预防结核分枝杆菌（Mtb）所致疾病和死亡的一种有前景的策略是接种疫苗。在本研究中，我们旨在评估一种由热休克蛋白X/分泌性蛋白EsxS融合蛋白、聚乳酸-羟基乙酸共聚物（PLGA）和1,2-二油酰基-3-三甲基铵丙烷（DOTAP）组成的多组分疫苗在BALB/c小鼠中引发抗Mtb免疫反应的能力。分别采用复乳溶剂蒸发法（w/o/w）和脂质膜水化法制备了含有融合蛋白和DOTAP佐剂的PLGA纳米颗粒（NPs）制剂。用不同制剂对BALB/c小鼠进行三次皮下免疫后，采用酶联免疫吸附测定（ELISA）技术检测脾细胞中干扰素-γ（IFN-γ）和白细胞介素-4（IL-4）细胞因子水平以及血清抗热休克蛋白X/分泌性蛋白EsxS IgG1和IgG2a滴度。本研究结果表明，与作为阳性对照的卡介苗（BCG）、热休克蛋白X/分泌性蛋白EsxS、热休克蛋白X/分泌性蛋白EsxS/DOTAP和PLGA/热休克蛋白X/分泌性蛋白EsxS制剂相比，PLGA/热休克蛋白X/分泌性蛋白EsxS/DOTAP制剂能够诱导更高水平的IFN-γ以及IgG1和IgG2a反应。我们的结果表明，PLGA纳米颗粒作为递送系统，DOTAP作为佐剂，在对BALB/c小鼠进行皮下免疫后，具有增强针对热休克蛋白X/分泌性蛋白EsxS抗原免疫反应的良好潜力。
